5-氨基酮戊酸介导的声动力疗法诱导巨噬细胞和泡沫细胞凋亡

Apoptosis of THP-1 Macrophages and Foam Cells Induced by Aminolevulinic Acidmediated Sonodynamic Therapy

目的探讨5-氨基酮戊酸(ALA)介导的声动力疗法(SDT)对巨噬细胞和泡沫细胞的作用,并进行比较。方法使用佛波酯诱导人急性单核白血病细胞(THP-1)成为巨噬细胞,再使用氧化型低密度脂蛋白诱导巨噬细胞成为泡沫细胞,使用这两种细胞进行实验研究。给予ALA共孵育后观察细胞内生成原卟啉Ⅸ(PpⅨ)的荧光强度;将两种细胞分别分为空白对照组和SDT组,并检测以下指标:DCFH-DA探针检测细胞内活性氧水平;线粒体膜电位探针(JC-1)检测细胞线粒体膜电位;Annexin V-PI染色后使用流式细胞仪检测细胞凋亡;酶活性探针检测细胞Caspase-9和Caspase-3的活性。结果给予ALA后3 h巨噬细胞和泡沫细胞均呈现较强的红色荧光,泡沫细胞荧光强度略高于巨噬细胞,但无统计学差异(P>0.05);SDT后巨噬细胞和泡沫细胞内活性氧分别增加1.84倍(P<0.01)和1.44倍(P<0.001);低线粒体膜电位的细胞分别增加1.96倍(P<0.001)和1.55倍(P<0.001);凋亡率分别升高1.36倍(P<0.001)和2.78倍(P<0.001);Caspase-9的酶活性分别升高1.34倍(P<0.05)和2.59倍(P<0.001);Caspase-3的酶活性分别升高2.97倍(P<0.01)和4.88倍(P<0.001)。结论 SDT通过超声作用于ALA-PpⅨ产生活性氧,损伤线粒体激活Caspase凋亡通路诱导巨噬细胞和泡沫细胞凋亡,对于泡沫细胞的作用更为显著,ALA介导的SDT可能成为减缓动脉粥样硬化斑块进程的一种新手段。

Aim To investigate and compare the effects of aminolevulinic acid（ ALA）-mediated sonodynamic therapy（ SDT） on macrophages and foam cells.Methods The THP-1 cells were differentiated into macrophages by adding phorbol-12-myristate-13-acetate,and then the macrophages were differentiated into foam cells by adding oxidized low density lipoprotein.The two kinds of cells were used to study.The protoporphyrin Ⅸ fluorescence intensity were observed after the cells incubated with ALA;The two kinds of cells were divided into control and SDT groups respectively,the following indicators were detected：reactive oxygen,mitochondrial membrane potential,cell apoptosis and the enzyme activity of caspase-9,3.Results Red PpⅨ fluorescence were observed in both macrophages and foam cells after incubation with ALA for three hours.The fluorescence of foam cells was stronger,but there was no statistical difference（ P〈0.05）;The reactive oxygen of macrophages and foam cells increased 1.84（ P〈0.01） and 1.44（ P〈0.001） times respectively after SDT;Cells with low mitochondrial membrane potential increased 1.96（ P〈0.001） and 1.55（ P〈0.001） times respectively;The apoptosis rate increased 1.36（ P〈0.001） and 2.78（ P〈0.001） times respectively;The enzyme activity of caspase-9increased 1.34（ P〈0.05） and 2.59（ P〈0.001） times;And the enzyme activity of caspase-3 increased 2.97（ P〈0.01） and4.88（ P〈0.001） times respectively.Conclusion ALA-mediated SDT had apoptotic effects on THP-1 macrophages and foam cells via generation reactive oxygen and damaging mitochondrial,activating caspase apoptosis pathway.It is more effective to foam cells.ALA-mediated SDT may be a potential treatment to attenuate progression of atherosclerotic plaque.